Lin Wei-Yu, Guven Ahmet, Juan Yung-Shun, Neuman Paul, Whitbeck Catherine, Chichester Paul, Kogan Barry, Levin Robert M, Mannikarottu Anita
Division of Urology, Department of Surgery, Chang Gung Memorial Hospital, Chia-Yi, Taiwan.
BJU Int. 2008 Mar;101(5):621-6. doi: 10.1111/j.1464-410X.2007.07389.x. Epub 2008 Jan 8.
To investigate the use of free-radical generation as a result of protein carbonylation and nitrotyrosination to characterize the level of bladder dysfunction after partial bladder outlet obstruction (PBOO) and reversal.
We surgically created PBOO in male New Zealand White rabbits; after 4 weeks of PBOO, one group of six rabbits was assessed, while the PBOO was relieved in two additional groups of six rabbits each that were assessed at 4 and 8 weeks after relieving the PBOO. Six sham-operated rabbits served as controls. Sedated rabbits were assessed by cystometry and the bladders were then removed for contractile, histological and molecular studies. Western blotting was used to determine the level of carbonylation and nitrotyrosination at the protein level.
The PBOO group had significant decreases in the contractile responses to field stimulation, ATP, carbachol and KCl. The responses to all forms of stimulation increased significantly at 4 weeks after reversal, and further increased to near normal levels by 8 weeks. Similarly, compliance and cystometric values also returned to near normal values after reversal. The hypertrophied smooth muscle of the obstructed bladders regressed to near-normal size. There was a significant increase in the level of carbonylation and nitrotyrosination after PBOO, and a progressive decrease in the 4-week reversal groups, nearing control values by 8 weeks.
Significantly increased carbonylation and nitrotyrosination levels after PBOO correlated with the severe dysfunction in the obstructed rabbits. Similarly, decreased levels of oxidation and nitration correlated with the functional recovery after reversal.
研究蛋白质羰基化和硝基化导致的自由基生成情况，以表征部分膀胱出口梗阻（PBOO）及解除梗阻后膀胱功能障碍的程度。
我们通过手术在雄性新西兰白兔身上制造PBOO；PBOO 4周后，对一组6只兔子进行评估，另外两组各6只兔子在解除PBOO后4周和8周时进行评估，每组兔子在评估时均已解除PBOO。6只假手术兔子作为对照。对镇静后的兔子进行膀胱测压评估，然后取出膀胱进行收缩性、组织学和分子研究。采用蛋白质印迹法在蛋白质水平测定羰基化和硝基化程度。
PBOO组对场刺激、ATP、卡巴胆碱和氯化钾的收缩反应显著降低。解除梗阻后4周，对所有形式刺激的反应均显著增加，并在8周时进一步增加至接近正常水平。同样，顺应性和膀胱测压值在解除梗阻后也恢复至接近正常水平。梗阻膀胱的肥厚平滑肌恢复至接近正常大小。PBOO后羰基化和硝基化水平显著升高，在4周解除梗阻的组中逐渐降低，至8周时接近对照值。
PBOO后显著升高的羰基化和硝基化水平与梗阻兔子的严重功能障碍相关。同样，氧化和硝化水平的降低与解除梗阻后的功能恢复相关。
